Journal
CURRENT PHARMACEUTICAL BIOTECHNOLOGY
Volume 16, Issue 2, Pages 162-178Publisher
BENTHAM SCIENCE PUBL LTD
DOI: 10.2174/1389201015666141113123910
Keywords
Box behnken design; identification; L-asparaginase production; plackett-burman design; 16S rRNA; Streptomyces parvus NEAE-95
Funding
- Science and Technology Development Fund (STDF), Egypt [4943]
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L-asparaginase is an effective anti-neoplastic agent used in chemotherapy of acute lymphoblastic leukemia. One hundred and thirty actinomycete isolates were isolated from several soil samples collected from different localities in Egypt. All these isolates were purified and evaluated for their ability to produce L-asparaginase activity. Among them, strain NEAE-95 was selected and identified as Streptomyces parvus strain NEAE-95 based on morphological, cultural, physiological characteristics and 16S rRNA sequence. The sequence was deposited in the NCBI GenBank database under accession number KJ200341. L-asparaginase production by Streptomyces parvus NEAE-95 was optimized in shake flask culture. The Plackett-Burman statistical design was used for initial screening of sixteen factors for their significance on L-asparaginase production. Among the variables screened, incubation time, L-asparagine and yeast extract had significant effects on L-asparaginase production. The levels of these significant variables and their interaction effects were optimized by Box-Behnken statistical design. As a result, the maximal L-asparaginase production was achieved at the following fermentation conditions: g/L (dextrose 2, starch 20, L-asparagine 14, KNO3 2, yeast extract 2, K2HPO4 2, MgSO4 center dot 7H(2)O 0.1, NaCl 0.1, FeSO4 center dot 7H(2)O 0.01), pH 7, temperature 30 degrees C, agitation speed 200 rpm, inoculum size 2%, v/v and incubation time 8 days.
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